Protection of hepatocytes from apoptosis by a novel substance from actinomycetes culture medium

A novel substance, #675, found from an Streptomyces sp. SM675 culture medium, dose-dependently stimulates the proliferation of human functional liver cell 4 (FLC4). When FLC4 cells were incubated under conditions without fetal bovine serum (FBS), typical features of apoptotic cell death such as shrinkage and nuclear condensation appeared; high molecular weight (HMW) DNA fragments were found; and caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were cleaved. When FLC4 cells were incubated with #675 and without FBS, the cells grew healthy, no HMW DNA fragments were found, and caspase-3 and PARP cleavage weakened, suggesting that #675 protects FLC4 cells from apoptosis induced by FBS-deprivation. The quantitative reverse-transcribed polymerase chain reaction did not show differences in PARP or Bcl-2 mRNA expression in FLC4 cells incubated with or without #675, indicating other genes may be involved in this anti-apoptosis effect. These results show that #675 enhances FLC4 proliferation via an apoptosis-inhibition pathway, implying potential pharmacological and clinical applications.     

Effects of different speeds of induction with sevoflurane on the EEG in Man

The effects of two kinds of induction speed of sevoflurane anesthesia on the EEG pattern were compared in the same individual using medical student volunteers: a first exposure of 4% was given, followed after full recovery, by incremental doses of 1, 2 and 4% successively, each being administered for 10min. The arterial blood level of the anesthetic was measured using gaschromatograph. The changes of EEG pattern during fast induction with 4% were not represented by the abbreviation of those observed during the slow induction with the incremental doses. The administration of 4% induced a sudden appearance of high voltage, rhythmic slow waves of 2–3Hz at 1–3min when the arterial blood anesthetic level increased maximally, which was then followed by a pattern of faster activities of 10–14Hz mixed with 5–8Hz slow waves. In contrast, the administration of incremental doses induced an increase in frequency and amplitude of EEG activities in the light plane, followed by their decreases in deeper planes. The final EEG patterns were identical for both these methods of induction. These findings confirmed our previous hypothesis that not only the arterial blood level of anesthetics but the rate of its increase are important factors determining the EEG pattern of anesthesia.(Avramov MN et al.: Effects of different speeds of induction with sevoflurane on the EEG in man. J Anesth 1: 1–7, 1987)     

The deubiquitinating enzyme Fam interacts with and stabilizes beta-catenin

BACKGROUND: In the ubiquitin-proteasome pathway, the ubiquitinated substrates either undergo degradation by the proteasome or stabilization through the action of the deubiquitinating enzyme. We have previously found that the deubiquitinating enzyme Fam is colocalized with AF-6, one of the effectors of the Ras small GTPase, at cell-cell contact sites in epithelial cells and interacts with AF-6 in vivo and in vitro. Fam has deubiquitinating activity in vitro and prevents the ubiquitination of AF-6 in intact cells. The degradation of beta-catenin, which accumulates at the cell-cell contact sites as a cadherin/catenin complex, is thought to be regulated by the ubiquitin-proteasome pathway. These observations prompted us to examine the possible Fam regulation of the stabilization of beta-catenin. RESULTS: We found that Fam interacted with beta-catenin both in vivo and in vitro. The Fam-binding site of beta-catenin mapped to the region close to the APC or Axin-binding site of beta-catenin. Over-expression of Fam in mouse L cells resulted in an elevation of beta-catenin levels and in an elongation of the half-life of beta-catenin. In these L cells, Fam was colocalized with beta-catenin at the dot-like structures in the cytoplasm. CONCLUSION: These results indicate that Fam interacts with and stabilizes beta-catenin in vivo, presumably through the deubiquitination of beta-catenin.     

Blood Gas Analysis Results and Serum Lactate Levels in Patients with Psychogenic Hyperventilation and Urinary Tract Infection with Suspected Sepsis: A Retrospective Comparative Study

Objective The prognosis differs considerably between patients with psychogenic hyperventilation syndrome (HVS) and those with urinary tract infection (UTI)-associated sepsis; however, the nonspecific symptoms and signs make the diagnosis and management difficult. We herein report the utility of a blood gas analysis for distinguishing HVS from UTI with suspected sepsis. Methods This single-center retrospective cohort study was conducted in a tertiary-care hospital in Japan. Patients ≥18 years old with a quick Sequential Organ Failure Assessment (qSOFA) score ≥2 and HVS or UTIs were included. The results of an arterial blood gas (ABG) or venous blood gas (VBG) analysis of the two groups were compared using the Mann-Whitney U test. We used a receiver-operating characteristic (ROC) curve analysis of the arterial pH and arterial PCO₂ to assess the ability of these analyses to distinguish HVS from UTI with suspected sepsis. Results A total of 64 patients with HVS (ABG, n=14; VBG, n=50) and 53 with UTI with suspected sepsis (ABG, n=35; VBG, n=18) were included. Patients with HVS had alkalemia and lower PCO₂ levels than patients with UTI with suspected sepsis, but the serum lactate levels were similar between the groups. The ROC analysis determined the pH cut-off value to be 7.509 (sensitivity: 0.91; specificity: 0.86) and the PCO₂ cut-off value to be 21.6 mmHg (sensitivity: 1.00; specificity: 0.64). Conclusion Elevated serum lactate levels alone cannot be used to differentiate between patients with HVS and those with UTI with suspected sepsis, but the degree of pH and PCO₂ abnormality can help with the differential diagnosis.     

Simple and rapid analysis of aristolochic acid contained in crude drugs and Kampo formulations with solid-phase extraction and HPLC photodiode-array detection

Simple and rapid analysis of aristolochic acid (AA) in crude drugs and Kampo extracts using a solid-phase extraction method and HPLC-PDA analysis was investigated. Extraction of AA from samples was accomplished by adding methanol containing 1% ammonia. The addition of ammonia ionized the AA of acidic substances so that they adhered to an acrylamide copolymer of a strong anion exchange resin (Sep-Pak QMA) coupled to diol silica easily. Furthermore, a mixture of acetonitrile–water–phosphoric acid (75:25:2, v/v) was effective in isolating AA from its carrier. Since almost all interfering peaks originating from contaminants in crude drugs and Kampo extract formulations could be removed, a satisfactory HPLC chromatogram of AA was obtained. A good result was also obtained when Aristolochiaceae and crude drugs containing AA were tested. Particularly in the case of the medicinal parts of Asarum, several interfering peaks and a ghost peak detected near the AA peak were eliminated. The AA contents of two Kampo extract formulations, tokishigyakukagoshuyushokyoto and ryutanshakanto, were calculated by HPLC analysis. The AA content (the sum of AA-I and AA-II) was 1.25–6.13 mg per daily dose. From an additional recovery experiment for Kampo formulations, high recovery rates of AA were obtained. Neither LC/MS nor special instrumentation was necessary. Our results suggest that this simple, quick, and sensitive analytical method to detect AA in crude drugs and Kampo extract formulations would be valuable in safety inspections of AA in crude drugs and their products.     

Advantages of Using Both Voxel- and Surface-based Morphometry in Cortical Morphology Analysis: A Review of Various Applications

Surface-based morphometry (SBM) is extremely useful for estimating the indices of cortical morphology, such as volume, thickness, area, and gyrification, whereas voxel-based morphometry (VBM) is a typical method of gray matter (GM) volumetry that includes cortex measurement. In cases where SBM is used to estimate cortical morphology, it remains controversial as to whether VBM should be used in addition to estimate GM volume. Therefore, this review has two main goals. First, we summarize the differences between the two methods regarding preprocessing, statistical analysis, and reliability. Second, we review studies that estimate cortical morphological changes using VBM and/or SBM and discuss whether using VBM in conjunction with SBM produces additional values. We found cases in which detection of morphological change in either VBM or SBM was superior, and others that showed equivalent performance between the two methods. Therefore, we concluded that using VBM and SBM together can help researchers and clinicians obtain a better understanding of normal neurobiological processes of the brain. Moreover, the use of both methods may improve the accuracy of the detection of morphological changes when comparing the data of patients and controls.In addition, we introduce two other recent methods as future directions for estimating cortical morphological changes: a multi-modal parcellation method using structural and functional images, and a synthetic segmentation method using multi-contrast images (such as T1- and proton density-weighted images).     

Osteocalcin fragment in bone matrix enhances osteoclast maturation at a late stage of osteoclast differentiation

Although 14-day-old mouse embryonic calvarial cells cultured in plastic culture dishes in the presence of 1,25-dihydroxyvitamin D₃[1,25-(OH)₂D₃] for 7 days could barely resorb bone slices, the same calvarial cells cultured with an ethylenediaminetetraacetic acid (EDTA) extract from bovine bone powder under the same conditions stimulated pit formation on bone slices in a dose-dependent manner. Therefore, the present study was conducted to purify and characterize this osteoclast maturation-inducing factor(s) from the bone matrix. The protein having osteoclast maturation-inducing activity in the EDTA extract was purified by gel filtration over Superdex 75 preparation grade and chromatography on hydroxyapatite, Mono Q, and C8 reversed-phase HPLC by monitoring the ability of the eluted fractions to elicit pit formation on bone slices. The molecular weight of the purified protein estimated by high-resolution polyacrylamide gel electrophoresis was 5.7kDa and 6.8kDa in the respective absence and presence of 2-mercaptoethanol. The sequence of the 30-amino-acid purified protein corresponded to the 7th to 36th residues of bovine osteocalcin. The osteocalcin fragment, missing the initial 6 residues at the N-terminal region, exhibited higher osteoclast maturation-inducing ability than bovine intact osteocalcin on a per weight basis. The osteocalcin fragment had no effect on the expression of receptor activator of nuclear factor (NF)-B ligand (RANKL) and osteoprotegerin (OPG) genes in calvarial cells, nor did it enhance the bone-resorbing activity of mature osteoclasts. When the osteocalcin fragment was added to late-stage (days 4–7) or to early-stage (days 0–3) cultures of calvarial cells pretreated with 1,25-(OH)₂D₃, its stimulatory effect was observed in the late-stage cultures rather than in the early-stage ones. In addition, the osteocalcin fragment directly enhanced the formation of osteoclasts with bone-resorbing ability from Mac-1⁺ c-Fms⁺ cells in the presence of macrophage colony-stimulating factor (MCSF) and RANKL. These results suggest that the osteocalcin fragment in bone matrix is involved in osteoclast maturation, especially at a late stage of osteoclast differentiation.     

Overview: Japanese guidelines for myocardial revascularization to treat stable ischemic heart disease 2012

Since its introduction in the early 1970s, coronary artery bypass grafting (CABG) has become an established surgical treatment for coronary artery disease (CAD). Percutaneous coronary intervention (PCI), first clinically applied in 1977, was promoted as an alternative to CABG during the mid-1980s. Along with the nationwide expansion of PCI, the ratio of PCI to CABG has exceeded 6–7:1. The Japanese Circulation Society (JCS) published “Guidelines on elective coronary intervention, including coronary artery bypass grafting (CABG), for ischemic heart disease in 2000” and “Guidelines on the selection of bypass conduits and operative procedures in coronary artery bypass grafting for ischemic heart disease”. The society intended to revise these guidelines in 2010 and to issue two new guidelines specific either to PCI or CABG. They also planned to issue joint guidelines for myocardial revascularization and PCI/CABG, which are primary indications for patients with stable CAD, especially with those with more complex left main trunk disease and/or multi-vessel disease. The scientific committee of JCS established the “Council for myocardial revascularization” that consisted of experts including interventional and non-interventional cardiologists, cardiac surgeons and physicians specialized in diabetes or nephrology selected by medical and surgical societies. After over 10 rounds of meetings, the Council prepared primary guidelines for myocardial revascularization to treat stable CAD, PCI/CABG. These guidelines consist of (1) Statements about myocardial revascularization, (2) Interpretation of the statements and (3) Indications for PCI/CABG including a table.